Figure 2.
Double-labeled proteins produced through Uaa mutagenesis and expressed protein ligation. (A) Incorporation of N-terminal thioamides through semisynthesis. (B) Incorporation of C-terminal thioamides through expressed protein ligation, where the expressed fragment is fused to a C-terminal intein. (C) Measurements of protein compaction in the presence of trimethylamine-N-oxide (TMAO) using α-synuclein containing a cyanophenylalanine (F*) and a thioamide (Thio-Ala, As or Thio-Phe, Fs) FRET pair; subscript indicates position in the α-synuclein sequence.